HPLC指纹图谱法研究峨眉黄连与三角叶黄连

目的建立三角叶黄连Coptis deltoidea和峨眉黄连Coptis omeinsis的根茎、根、叶的高效液相指纹图谱。方法采用HPLC-DAD法测定峨眉黄连、三角叶黄连及黄连的根茎、根和叶中小檗碱的含量及各部位的指纹图谱,用指纹图谱相似度评价方法综合比较黄连药材的差异。结果从化学组分及各组分含量的相似度综合比较,同一分布区域内的3种黄连中,峨眉黄连与三角叶黄连更相似,同为雅连使用有一定的依据。结论所用方法简便、重复性好,可用于黄连属植物的化学组分与种间鉴别的比较依据。     

岩黄连注射液的高效液相色谱质谱联用指纹图谱研究

目的建立岩黄连注射液的指纹图谱。方法色谱部分采用高效液相色谱梯度洗脱法，色谱柱为Kromasil 100-10C₁₈柱(250 mm×4.6 mm ID,5 μm)；流动相为乙腈和水；流速为0.5 mL·min^-1；在254 nm波长下进行分析。质谱部分采用电喷雾离子阱质谱分析，正离子扫描方式。结果通过LC/MS对其中的11种主要成分进行了指认，LC/UV对其中的2种成分进行了定量评价。结论所建指纹图谱能体现岩黄连注射液的特点，相似度评价和方法学考察结果良好，可用于质量控制及定量测定。     

岩黄连药材HPLC指纹图谱的研究

目的:采用 HPLC 法建立岩黄连药材指纹图谱。方法:应用 Shim—pack CLC—ODS 色谱柱(6.0 mm×150 mm,5μm),乙腈-缓冲溶液[0.05 mol·L~(-1)庚烷磺酸钠-0.05 mol·L~(-1)磷酸二氢钾(1:1),含0.2%乙二胺,用磷酸调 pH 3.0]梯度洗脱:0～130 min,流动相比例20:80→27:73;130～140 min,比例27:73→20:80;140～160 min,比例20:80。流速为1 mL·min~(-1),检测波长285 nm,柱温:室温。以中药色谱指纹图谱相似度评价系统的操作规范(版本2004 A)软件计算。结果:10批药材的相似度在0.765～0.989,确定了8个共有峰,建立了岩黄连药材 HPLC 指纹图谱。结论:本法可作为岩黄连药材的质量控制方法。     

黄连解毒汤饮片汤剂和颗粒汤剂的指纹图谱比较

目的:用指纹图谱对黄连解毒汤传统饮片汤剂和现代颗粒汤剂进行比较.方法:采用RP-HPLC梯度洗脱进行分析,色谱柱:Diamonsil(TM)-C18柱(250 mm×4.6 mm);流动相:水-甲醇-0.05%磷酸(梯度洗脱);检测波长:254 nm;柱温:35℃.结果:所建立分析方法得到的HPLC色谱图有较好重现性.结论:不同厂家饮片汤剂、颗粒汤剂指纹图谱存在一定的差异.     

黄连厚朴乙醇分提和合提提取物的指纹图谱比较

目的 以指纹图谱方式，研究黄连和厚朴乙醇合提和分提提取物中化学成分的差异。方法 采用常规制备方法得黄连和厚朴醇提合提和分提的提取物，采用HPLC考察并确定最佳的检测波长，最佳色谱柱等色谱条件，并采用最佳色谱条件对黄连厚朴配伍不同制备方法的提取物进行测定，比较不同提取物的指纹图谱的差异。结果 不同提取物中化学成分组成存在差异，且提取物中主要化学成分的提取率也存在差异。结论 黄连厚朴醇提合提和醇提分提提取物中主要化学成分组成存在差异。     

黄连厚朴乙醇分提和合提提取物的指纹图谱比较

目的以指纹图谱方式,研究黄连和厚朴乙醇合提和分提提取物中化学成分的差异。方法采用常规制备方法得黄连和厚朴醇提合提和分提的提取物,采用HPLC考察并确定最佳的检测波长,最佳色谱柱等色谱条件,并采用最佳色谱条件对黄连厚朴配伍不同制备方法的提取物进行测定,比较不同提取物的指纹图谱的差异。结果不同提取物中化学成分组成存在差异,且提取物中主要化学成分的提取率也存在差异。结论黄连厚朴醇提合提和醇提分提提取物中主要化学成分组成存在差异。     

复方黄连胶囊的稳定性研究

目的考察复方黄连胶囊的稳定性。方法采用留样观察,在室温下,分别于0、1、2、3、6、12、18、24个月对样品的外观、性状进行观察,对盐酸小檗碱、葛根素、熊果酸进行定性鉴别,并对盐酸小檗碱的含量进行测定。结果按本品最佳制备工艺条件制成的3批中试产品的外观、性状、定性鉴别、含量测定结果均符合规定。结论复方黄连胶囊稳定性符合标准。     

配伍对黄连解毒汤及其拆方HPLC-DAD指纹图谱的影响

目的：研究不同配伍对黄连解毒汤及其拆方中主要特征指纹峰的影响。方法：采用HPLC方法分析全方及不同拆方样品,色谱柱为Diamonsil C18柱（250mm×4．6mm,5μm）;以乙腈-0．1％冰乙酸为流动相进行线性梯度洗脱;流速：1．0ml·min^-1;柱温：40℃;UV检测波长：260nm。结果：全方煎剂的特征指纹峰基本为各药味特征峰的加和,不同配伍对黄连解毒汤中主要特征峰有不同影响,但未产生明显的新特征峰。结论：本研究为黄连解毒汤的配伍规律及物质基础研究提供了一定参考。     

酒蒸黄连HPLC指纹图谱及其与不同黄连炮制品的比较研究

目的:建立酒蒸黄连的HPLC指纹图谱,并比较不同黄连炮制品的差异.方法:采用HPLC法测定了酒蒸黄连等不同炮制品样品,运用主成分分析和相似度评价方法分析其结果.结果:建立了酒蒸黄连的HPLC指纹图谱,主成分分析将七批酒蒸黄连样品聚集在一起,并与其它炮制品样品明显分离.结论:酒蒸黄连与姜黄连、萸黄连、酒黄连等炮制品化学成分有一定的差异.本法稳定、准确、重复性好,为酒蒸黄连的质量标准研究提供了科学依据.     

基于指纹图谱的黄连解毒汤8种指标成分含量测定及多元统计分析

目的:建立黄连解毒汤的指纹图谱,测定其中8种成分的含量,并进行正交偏最小二乘判别法(OPLS-DA)分析。方法:采用HPLC法,色谱柱为Sepax Bio-C₁₈,流动相为甲醇-0.05%磷酸溶液(梯度洗脱),检测波长为238 nm,柱温为35℃,流速为0.6 mL·min^-1。采用《中药色谱指纹图谱相似度评价系统》2012年版进行相似度评价,确定共有峰,并测定其中8种成分的含量;采用SIMCA 14.0软件进行OPLS-DA分析,并利用分析变量重要性投影值(VIP)筛选影响其质量的标志物。结果:10批黄连解毒汤共有18个共有峰,与对照指纹图谱的相似度均>0.98;共指认了8个色谱峰并分别测定了含量,各成分在考察的范围内线性良好,r值在0.999 1～0.999 9之间,平均加样回收率为97.89%～100.10%(RSD为1.19%～2.36%,n=6)。OPLS-DA分析结果显示,盐酸小檗碱、栀子苷、黄芩苷是影响黄连解毒汤质量的主要成分物质。结论:结合多元统计分析,建立的指纹图谱和含量测定方法准确、稳定、可行,可用于黄连解毒汤的质量...     

《中华人民共和国药典》与美、英、日3国《药典》含量均匀度检查法的比较

目的比较《中华人民共和国药典》与美、英、日3国《药典》含量均匀度检查法的异同,尝试建立新的含量均匀度检查法。方法采用计算机模拟随机抽样方法,绘制《中华人民共和国药典》含量均匀度检查法和ICH方法的抽样特性曲线,计算统计特性参数。通过比较2种方法的特征曲线与抽样特性参数,分析2种方法的优点和产生原因,提出ICH改进方法并计算ICH改进方法的抽样特性参数。结果《中华人民共和国药典》含量均匀度检查方法I、CH方法及ICH改进方法的批允许废品率、复试率、平均样本容量和争议区分别为9.4%、27.4%、15.5和15.0%;5.5%、69.6%、23.9和11.0%;5.5%、40.5%、18.1和11.0%。结论ICH改进方法优于ICH方法。     

香连制剂中黄连的质量分析

目的:通过对127批香连制剂样品进行标准检验及探索性研究,对该制剂质量标准中黄连的检测项进行分析和评价,探索香连制剂中黄连新的质量控制方法。方法:标准检验依据《中国药典》2010年版一部(片剂和丸剂)和国家药品标准新药转正标准第65册(胶囊剂),检验项目包括TLC鉴别及HPLC含量测定。探索性研究采用薄层制备、HPLC-DAD、HPLC-MS等检测方法,对香连胶囊中未知成分进行了分析鉴定;采用一测多评技术(QAMS)测定4种黄连生物碱含量。结果:按现行标准,合格率100%;与对照品比对,确认香连胶囊样品中的未知成分为小檗红碱;较多厂家的香连制剂中小檗碱的含量远高于其他黄连生物碱,致使制剂与黄连药材中生物碱的含量比例差异较大,存在非法添加问题。结论:探索性研究为进一步修订标准及有效地控制该制剂质量提供参考依据。     

益眠达片的稳定性试验

目的 根据益眠达片制剂质量标准,对益眠达片的稳定性进行考察.方法 采用加速试验和长期试验,考察益眠达片的鉴别、检查、性状、崩解时限、重量差异、微生物限度、五味子醇甲含量等检测项的变化.结果 益眠达片供试品经加速试验6个月考察,检测项均无明显改变;供试品经长期试验18个月,在性状、鉴别、崩解时限方面都没有显著性变化.但是,在微生物检查方面,供试品在放置到第18个月时,有2批供试品的细菌数每1 g超过10 000 efu,不符合微生物限度检查法项下的规定.结论 根据试验数据,为保证益眠达片在使用过程中的安全,暂定益眠达片有效期为12个月.     

MDR1 bicistronic vectors: analysis of selection stringency, amplified gene expression, and vector stability in cell lines

The human multidrug resistance-1 gene (MDR1) is a dominant selectable and amplifiable marker in mammalian tissue culture cells. MDR1 is also being investigated as a gene therapy tool, both to protect normal cells against chemotherapy-related toxicity and to serve as an in vivo selectable marker for the overexpression of non-selectable therapeutic genes. The success of these strategies will depend on whether MDR1 expression can be sustained at levels high enough to confer a survival advantage on target cells. However, the MDR1 selection system is quite stringent, requiring high gene expression for transduced cells to survive in the presence of drug. The current report is a detailed molecular analysis of MDR1 selection stringency compared with the common neo selectable marker. A bicistronic vector encoding MDR1 and neo genes linked through an internal ribosome entry site was transferred into NIH 3T3 mouse fibroblasts and K562 human leukemia cells; cells were then exposed to colchicine (to select for MDR1 expression) or to G418 (to select for neo expression). Surviving populations and individual clones of cells were analyzed for expression levels of MDR1 and neo gene products; resistance to colchicine, paclitaxel, and G418; level and integrity of bicistronic mRNA; and structural integrity, integration number, and copy number of vector DNA. These studies provide direct evidence that colchicine selection is more stringent than G418 selection; that increased selection pressure with colchicine leads to increased gene expression; that increased gene expression can be accommodated primarily by gene amplification, even within an individual transduced clone and starting from a single-copy proviral integration event; and that the clonal diversity of a transduced population of cells is influenced significantly by the stringency of selection. Taken together, these results have important implications for the potential utility of MDR1 as a selectable marker and as a gene therapy tool in hematopoietic cells.     

Chemical stability of lipid excipients in SLN-production of test formulations, characterisation and short-term stability

The study investigates the chemical stability of lipids used as excipients in the production of solid lipid nanoparticles (SLN). A total of 17 SLN formulations was produced using different lipids. Most of the formulations were produced using identical binary surfactant mixtures and concentrations to study the effect of the chemical nature of the lipid on its stability in SLN. In some formulations surfactants were exchanged to study the contribution of the surfactant. The particles were characterised by photon correlation spectroscopy, laser diffractometry, zeta potential determination and differential scanning calorimetry, the latter to assess potential effects of lipid crystallinity and modifications on lipid stability. Lipid analysis was performed by gas chromatography using a sampling preparation and analysis procedure especially developed for SLN. This short-term study provides primarily information about the stability of the lipid under production conditions, that means high pressure homogenisation (cavitation) at high temperature. No degradation products couldbe detected for all lipids, the production process itself did not impair excipient stability.     

Content uniformity test of tablets by nonaqueous titration method

For the Content uniformity test (CUT) of tablets where the active drug behaves in a nonaqueous medium as a base a very simple method was developed. This method is based on the extraction of the active drug from the tablet into anhydrous acetic acid and direct titration of the solute with HClO4 (0.1 or 0.05 mol/l solution in anhydrous acetic acid). The course of the titration is followed potentiometrically with a glass and calomel electrode coupled and recorded automatically with a suitable registration potentiometer. The method was successfully applied to samples of commercial tablets containing the following drugs: codeine phosphate, embramine, ephedrine, ethylmorphine, pyridoxine, thiamine and tolazoline, all as hydrochlorides. It can be used also for several other drugs.     

The reliability and stability of the Mortimer-Filkins test.

The Mortimer-Filkins test has been used widely as an instrument for detecting problem drinkers among drink-driving offenders. While extensive psychometric testing has been undertaken by the developers of the test, few independent validation studies have been conducted, and few studies have used the Mortimer-Filkins test with general populations. The present study investigated the test-retest and internal-consistency reliability of the instrument, and the stability of problem drinking, as measured by the instrument. The test was administered to moderate and heavy drinkers at an industrial workplace on three occasions. The results indicated that the Mortimer-Filkins test has high test-retest and internal-consistency reliability, and problem drinking, as measured by the test, appears to be a stable characteristic across time.